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L-selectin mediates neutrophil rolling in inflamed venules through sialyl
LewisX-dependent and -independent recognition pathways
UH von Andrian, JD Chambers, EL Berg, SA Michie, DA Brown, D Karolak, L Ramezani, EM Berger, KE Arfors and EC Butcher
La Jolla Institute for Experimental Medicine, CA.
The glycoprotein (GP) L-selectin initiates adhesive interactions between
leukocytes and endothelial cells (EC). It functions as a lymphocyte-lectin
homing receptor recognizing carbohydrate determinants of the peripheral
lymph node addressing on high endothelial venules. It also mediates
neutrophil rolling, the earliest interaction of neutrophils with acutely
inflamed venules. Neutrophil L-selectin presents sialyl-LewisX (sLe(X)) as
a ligand to P- and E-selectin in vitro, and we have proposed that this is a
major mechanism of L- selectin-mediated rolling in vivo. In contrast, the
contribution of neutrophil L-selectin as a receptor protein recognizing one
(or more) ligand(s) on inflamed EC is unclear. To address this question, an
sLe(X)-negative murine pre-B cell line, L1-2, that can neither bind
vascular selectins nor roll in inflamed rabbit venules, was transfected
with human L-selectin cDNA. L-selectin expression in stable transfectants
was sufficient to confer significant rolling in vivo. Rolling was
unaffected by neuraminidase treatment but completely blocked by
anti-L-selectin monoclonal antibody (MoAb) DREG-56. Thus, L- selectin can
initiate leukocyte interactions with EC determinants potentially through
recognition of endothelial carbohydrates. In contrast, when human
neutrophils were tested, rolling was reduced, but not abolished, by MoAb
DREG-56. Likewise, treatment with neuraminidase or anti-sLe(X) MoAbs
decreased, but did not abrogate, neutrophil rolling, consistent with
residual EC recognition via L-selectin. Combination of MoAb DREG-56 and
neuraminidase resulted in almost complete loss of rolling, as did removal
of glycosylated L-selectin by chymotrypsin. Together with the demonstrable
rolling of L-selectin transfectants, our results support the concept of a
bidirectional interaction between L-selectin bearing sLe(X) on neutrophils
and activated EC in vivo. These findings also suggest that L-selectin may
mediate rolling of lymphocytes that lack carbohydrate ligands for E- or
P-selectin, although probably less efficiently than through bidirectional
recognition.
Volume 82,
Issue 1,
pp. 182-191,
07/01/1993
Copyright © 1993 by The American Society of Hematology
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